NAMSA, Cheshire, UK.
Natrox Wound Care (Inotec AMD Ltd.), Cambridge, UK.
J Wound Care. 2024 Sep 2;33(9):702-707. doi: 10.12968/jowc.2024.0213.
The effect of continuous topical oxygen therapy (cTOT) on biofilm gene transcription profiles following inoculation onto porcine skin, using a customised molecular assay was determined.
Sterilised porcine skin explants were inoculated with in triplicate: 0 hours as negative control; 24 hours cTOT device on; 24 hours cTOT device off. The oxygen delivery system of the cTOT device was applied to the inoculated tissue and covered with a semi-occlusive dressing. All samples were incubated at 37±2°C for 24 hours, with the 0 hours negative control inoculated porcine skin samples recovered immediately. Planktonic suspensions and porcine skin biopsy samples were taken at 0 hours and 24 hours. Samples were processed and quantifiably assessed using gene specific reverse transcription-quantitative polymerase chain reaction assays for a panel of eight genes (16S, pelA, pslA, rsaL, pcrV, pscQ, acpP, cbrA) associated with biofilm formation, quorum sensing, protein secretion/translocation and metabolism.
Transcriptional upregulation of pelA, pcrV and acpP, responsible for intracellular adhesion, needletip protein production for type-3 secretion systems and fatty acid synthesis during proliferation, respectively, was observed when the cTOT device was switched on compared to when the device was switched off. Data suggest increased metabolic activity within bacterial cells following cTOT treatment.
cTOT is an adjunctive therapy that supports faster healing and pain reduction in non-healing hypoxic wounds. Oxygen has previously been shown to increase susceptibility of biofilms to antibiotics through enhancing metabolism. Observed gene expression changes highlighted the impact of cTOT on biofilms, potentially influencing antimicrobial treatment success in wounds. Further in vitro and clinical investigations are warranted.
使用定制的分子检测方法,确定连续局部氧疗(cTOT)对猪皮接种后生物膜基因转录谱的影响。
将无菌猪皮样本以一式三份接种:阴性对照(0 小时);cTOT 装置开启 24 小时;cTOT 装置关闭 24 小时。将 cTOT 装置的供氧系统应用于接种组织,并覆盖半封闭敷料。所有样本在 37±2°C 下孵育 24 小时,立即回收 0 小时阴性对照接种的猪皮样本。在 0 小时和 24 小时时取浮游菌悬液和猪皮活检样本。通过基因特异性逆转录定量聚合酶链反应(RT-qPCR)检测 8 个与生物膜形成、群体感应、蛋白分泌/转运和代谢相关的基因(16S、pelA、pslA、rsaL、pcrV、pscQ、acpP、cbrA)的表达水平,对样本进行处理和定量评估。
与关闭 cTOT 装置相比,当打开 cTOT 装置时,观察到 pelA、pcrV 和 acpP 基因的转录上调,这些基因分别负责细胞内黏附、III 型分泌系统的针状蛋白产生和增殖过程中的脂肪酸合成。这表明 cTOT 处理后细菌细胞的代谢活性增加。
cTOT 是一种辅助治疗方法,可支持非愈合缺氧性伤口更快愈合和减轻疼痛。先前的研究表明,氧气通过增强代谢作用增加生物膜对抗生素的敏感性。观察到的基因表达变化突出了 cTOT 对生物膜的影响,这可能影响伤口的抗菌治疗效果。需要进一步进行体外和临床研究。